[Effects of Zhibai Dihuang Decoction on mitochondrial cytochrome oxidase in the spermatogenic cells of rats with ureaplasma urealyticum infection].

OBJECTIVE: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (ï¼»1.03 ± 0.09ï¼½ vs ï¼»0.07 ± 0.03ï¼½ %, P<0.01), grade b sperm (ï¼»2.07 ± 0.52ï¼½ vs ï¼»0.35 ± 0.13ï¼½ %, P<0.01), straight line velocity (VSL) (ï¼»10.95 ± 0.98ï¼½ vs ï¼»6.78 ± 1.05ï¼½ µm/s, P<0.01), curvilinear velocity (VCL) (ï¼»42.03 ± 1.35ï¼½ vs ï¼»38.10 ± 7.65ï¼½ µm/s, P>0.05), average path velocity (VAP) (ï¼»16.22 ± 1.52ï¼½ vs ï¼»10.05 ± 1.80ï¼½ µm/s, P<0.01), and the mRNA expressions of COX Ⅰ (ï¼»2.25 ± 0.24ï¼½ vs ï¼»0.93 ± 0.10ï¼½ %, P<0.01) and Ⅱ (ï¼»6.72 ± 0.37ï¼½ vs ï¼»2.95 ± 0.78ï¼½ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ï¼»1.11 ± 0.30ï¼½ and ï¼»0.60 ± 0.19ï¼½%; grade b sperm: ï¼»2.40 ± 0.59ï¼½ and ï¼»1.32 ± 0.27ï¼½ %; VSL: ï¼»12.11 ± 1.62ï¼½ and ï¼»11.47 ± 1.21ï¼½ µm/s; VCL: ï¼»54.30 ± 2.35ï¼½ and ï¼»45.75 ± 1.64ï¼½ µm/s; VAP ï¼»18.40 ± 1.27ï¼½ and ï¼»16.69 ± 1.02ï¼½ µm/s; expression of COXⅠ mRNA: ï¼»1.86 ± 0.30ï¼½ and ï¼»1.74 ± 0.17ï¼½ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS: UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.

[Effect of Zhibai Dihuang Decoction on Expressions of TRPV1 and TRPV5 in Spermatogenic Cells of UU-infected Rats].

OBJECTIVE: To observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD. METHODS: Totally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot. RESULTS: The UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01). CONCLUSION: ZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.